CEEPC/IPM/CMSC - Abstrakt prezentace

(CEEPC/IPM/CMSC 2022 - ThO-01)
Identification of potential cell-surface targets and druggable enzymes in human pheochromocytoma and paraganglioma using classical and membrane-targeting proteomic approaches

Ondřej Vít 1 *, Zdeněk Musil 2,3, Igor Hartmann 4,5, Zdeněk Fryšák 6,5, Karel Pacák 7, Jiří Petrák 1

  1. Charles University, 1st Faculty of Medicine, BIOCEV, Vestec
  2. Charles University, 1st Faculty of Medicine, Institute of Biology and Medical Genetics
  3. General University Hospital in Prague
  4. University Hospital Olomouc, Department of Urology
  5. Palacký University, Faculty of Medicine and Dentistry, Olomouc
  6. University Hospital Olomouc, Department of Internal Medicine III
  7. Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, Bethesda, USA

Abstrakt

Integral membrane proteins (IMPs) represent optimal drug targets but are under-represented in standard proteomic analyses due to their amphipathy, lack of trypsin cleavage sites, and low expression levels. To identify tumor-upregualted IMPs and druggable enzymes in rare neuroendocrine tumors – pheochromocytoma and paraganglioma (PPGL), we combined three approaches: 1) hpTC method, where the identification of IMPs is based on their hydrophobic alpha-helices, isolated by proteolytic shaving and re-cleaved with CNBr; 2) two glycocapture methods - lectin entrapment on ultrafilters (N-glyco-FASP), and solid-phase enrichment with hydrazide chemistry (SPEG); and 3) the classical detergent-trypsin approach.
We focused mainly on the high-risk tumors belonging to the so-called cluster 1, characterized by mutations in genes related to citric acid cycle and hypoxia, such as SDHB, VHL, and EPAS1). The classical trypsin-based proteomic approach pointed us toward upregulated soluble druggable enzymes (autotaxin, SHMT2, and Arginase 2) and upregulated cell surface IMPs (including CD146 and CD171). The glycopeptide enrichment approach provided additional potential cell surface targets (CD39), and the hpTC method provided additional IMP targets (e.g., anoctamin-1).
The above-mentioned IMPs and soluble enzymes have all been previously shown to be upregulated in several human cancers and to affect tumor progression directly. Their marked upregulation was confirmed by specific antibodies. Together, this makes these molecules promising candidates for drug targets and/or proteins enabling sensitive tumor imaging.

* Korespondující autor: ondrvit@gmail.com


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